Fibrin sealants or adhesives comprising a hyaluronic acid derivative material

ABSTRACT

An especially useful fibrin glue composition comprises a biocompatible, bioabsorbable hyaluronic acid derivative material, upon which are applied or chemically bonded fibrinogen and thrombin, along with other optional constituents, such as additional coagulation factors, anti-fibrinolytics, stabilizers and biologically active substances. The fibrinogen, thrombin and other components can take the form of a dry preparation, an aqueous or nonaqueous preparation, or as a combination thereof. Such a fibrin glue composition can be placed directly on a wound site and is fully reabsorbed into the body.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Application Ser. No.09/554,745, filed Aug. 14, 2000, now U.S. Pat. No. 6,503,527 as anational stage application under 35 U.S.C. §371 based on InternationalApplication No. PCT/US98/24605, filed Nov. 17, 1998, which claims thebenefit of the priority date under 35 U.S.C. §119 of U.S. ProvisionalApplication No. 60/065,884, filed Nov. 17, 1997.

BACKGROUND OF THE INVENTION

Fibrin glues, also called fibrin adhesives or sealants, are known foruses in a medical context. Generally, a fibrin glue is obtained byadmixing fibrinogen and thrombin containing components. The componentsare mixed, allowing the thrombin to convert the fibrinogen to fibrinmonomers. A number of methods for the production of fibrin glues areknown, as illustrated by: Thompson et al., “Fibrin Glue: A Review of ItsPreparation, Efficacy, and Adverse Effects as a Topical Hemostat,” DrugIntelligence and Clinical Pharmacy 22: 946-52 (1988); Brennan, “FibrinGlue,” Blood Reviews 5: 240-44 (1991); Stechison, “Rapid PolymerizingFibrin Glue from Autologous or Single Donor Blood: Preparation andIndications,” J. Neurosurgery 76: 626-28 (1992); and Toma et al.“Autologous Fibnn Glue in the Repair of Dural Defects in CraniofacialResections,” J. Laryngology and Otology 106: 356-57 (1992). (Therespective contents of publications cited in this description hereby areincorporated by reference.)

The fibrinogen component of the composition can be obtained byconventional methodology. Examples of such methods includecentrifugation, cryoprecipitation and precipitation using polyethyleneglycol, ether, ethanol, glycine or ammonium sulfate. Methods ofobtaining suitable fibrinogen are disclosed, for example, by Brennan,“Fibrin Glue,” Blood Reviews 5: 240-244 (1991). Further examples offibrin components are disclosed in U.S. Pat. Nos. 5,290,918 and5,395,923.

The thrombin component of the composition is also well known in the artand can be obtained by conventional methods, including recombinantmethods. Bovine and human derived thrombins are illustrative ofavailable thrombins well known in the art.

Application of the fibrin glue can be accomplished in a number of waysknown in the art. In one method, the admixture is drawn into a syringeand ejected via an appropriate sized needle. In another method a doublebarrel syringe is used. Other conventional techniques employ a microdropdelivery system, a spray application via a multi-channel catheter whichis fixed to a pressurized gas source, or a carrier, such as collagenfleece, dura, or a graft. Additionally, a number of special applicatorsare commercially available.

Numerous uses for fibrin glues are known. Fibrin glues are used in avariety of medical procedures as hemostatic agents, sealants andadhesives. For example, see Chisholm et al., “Fibrin Sealant as a Plugfor the Post Liver Biopsy Needle Track,” Clinical Radiology 40: 627-28(1989); Toma et al., “Autologous Fibrin Glue in the Repair of DuralDefects in Craniofacial Resections,” J. Latyngology and Otology 106:356-57 (1992); Kjaergard et al., “Autologous Fibrin Glue Preparation andClinical Use in Thoracic Surgery,” Eur. J. Cardio-Thorc. Surg. 6: 52-54(1992); Thompson et al., “Fibrin Glue: A Review of Its Preparation,Efficacy, and Adverse Effects as a Topical Hemostat,” Drug Intelligenceand Clinical Pharmacy 22: 946-52 (1988); Brennan, “Fibrin Glue,” BloodReviews 5: 240-44 (1991).

Available fibrin glues, however, have a number of significantdisadvantages. The fibrinogen and thrombin components must be mixed justprior to use. Admixing too early can result in clotting before it can beapplied. Thus, medical professionals are forced to divert theirattention for a significant amount of time to prepare the fibrin glue.Early clotting also causes problems in application, such as clogging inthe needle or applicator. Additionally, preparation of the fibrin gluecan involve complex and time-consuming efforts to establish a workablemix of the fibrinogen and thrombin components. Still further, availablefibrin glues may insufficiently adhere to the wound, or may provideinadequate strength to the wound during the healing process.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to a fibrin glue composition comprising abiocompatible and bioabsorbable material, i.e., a biomaterial, of ahyaluronic acid (HA) or a hyaluronic acid derivative. The material maybe either nonfilamentous (a film) or a woven or nonwoven fabric. Thefibrin glues of the present invention have fibrinogen and thrombinapplied to or chemically bonded to the HA or HA derivative material.Additional elements also can be applied to the material. Exemplary ofthese additional elements are further coagulation factors,anti-fibrinolytics, stabilizers and biologically active substances.

The fibrinogen, thrombin and, optionally, other elements can be appliedto the hyaluronic acid derivative film as a dry preparation, as anaqueous or nonaqueous preparation, or as a combination thereof.

BRIEF DESCRIPTION OF THE DRAWING(S)

Not Applicable

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a fibrin glue composition comprised ofa biocompatible, bioabsorbable hyaluronic acid derivative material,having applied thereto fibrinogen and thrombin. A composition accordingto the present invention also can contain additional blood coagulationfactors, stabilizers, fibrinolysis inhibitors and/or biologically activesubstances. Factor II and Factor XIII are examples of additionalcoagulation factors which are suitable for use with the presentcompositions. Anti-fibrinolytics, such as aprotinin and ε-aminocaproicacid, can also be used. A wide range of biologically active substancesare suitable for use with the present invention, including antibiotics,chemotherapeutics, fibroblastic growth factors, angiogenic growthfactors, anti-angiogenic growth factors, anti-neoplastic agents, cellgrowth factors and differentiating agents. Additionally, a suitablesource can be added for calcium ions, such as calcium salts.

When placed on a wound site and activated, the compositions of thepresent invention function as a fibrin glue. In contrast to conventionalfibrin glues, however, the inventive compositions do not require complexmixing of fibrinogen and thrombin components immediately prior to useand do not require special applicators.

The compositions of the present invention can be put to a wide range ofsuitable medical and surgical uses. The compositions can be used inhemostasis applications, as sealants and as adhesives. The compositionsof the present invention have a number of surgical uses. Incardiovascular surgery, they can be used as a hemostatic, for example,with needle holes, suture lines, diffuse and nonspecific bleeding,friable tissue bleeding, aortic dissections, ventricular ruptures, andfistulas. In otorhinolaryngology (ear, nose and throat, ENT) surgery,they can be used in facial nerve grafts, closure of dural leaks, nasalseptal surgeries, and post tonsillectomy hemorrhage. In neurosurgery,they can be used to prevent cerebral spinal fluid (CSF) leakage,peripheral nerve repair, and to anchor dural patches. In plasticsurgery, they can be used in a number of procedures relating to skingrafts, including to fix grafts, control oozing and control bleeding. Inthoracic surgery, they can be used, for example, in the treatment ofpneumothorax and pulmonary leaks. The compounds of the present inventionalso have a number of other surgical uses; Illustrative examplesinclude, sealing biopsy needle tracks, liver and splenic lacerations,lymphatic fluid leaks, organ resectioning, seroma and hematomaprevention, and gastrointestinal bleeding. The compositions of thepresent invention also can be used as a local delivery vehicle for thedelivery antibiotics or other biologically active substances to theapplication site. The compositions of the present invention also mayserve as a surgical adhesion barrier. Other uses are known in the art orwill be apparent to the skilled artisan.

In one embodiment, the present invention utilizes total or partialcross-linked hyaluronic acid derivatives. These hyaluronic acidderivative compositions can be made in the form of biocompatible andbioabsorbable films. These compositions and methods of making thesecompositions are disclosed in numerous publications, including U.S. Pat.No. 4,957,744 (Cross-linked Esters of Hyaluronic Acid), U.S. Pat. No.4,851,521 (Esters of Hyaluronic Acid), U.S. Pat. No. 5,202,431 (PartialEsters of Hyaluronic Acid), and U.S. Pat. No. 5,336,767 (Total orPartial Esters of Hyaluronic Acid). The specific hyaluronic compositioncan be selected by reference to desired properties, such ashydrophobicity and reabsorption time. The hyaluronic acid derivativesthus employed are formed into films as described in the art. These filmscan be shaped according to their intended use.

In another embodiment, the present invention utilizes esters of HA.These HA esters typically are formed by treating quaternary ammoniumsalt of HA with an esterifying agent in a suitable aprotic solvent.Esterification may be carried out using a number of different classes ofalcohols such as aliphatic, aryliphatic, cycloaliphatic andheterocyclic. Thus, a number of different derivatives can besynthesized. These derivatives also have a wide range of physicochemicalproperties. HA esters suitable for use in the present invention aredescribed, for example, by Benedetti et al., “Chemical Modification ofHyaluronan,” in Novel Biomaterials Based in Hyaluronic Acid and ItsDerivatives, Proceedings of a Workshop Held and the Annual Meeting ofthe European Society for Biomaterials (Pisa, Sep. 10, 1994) (hereafter“1994 Novel Biomaterials Proceedings”). See also European Patent 216453and Benedetti, “New Biomaterials from Hyaluronic Acid,” Medical DeviceTechnology (November 1994), pages 32-37.

These esters may be formed into various materials, such as fibers,membranes, sponges, fleece-like materials and threads. The fibers may beutilized to produce various fabrics, including woven or nonwovenmaterials. The membranes also can be made into perforated membranes.Materials comprised of HA esters are detailed by Hellstrom et al.,“Endogenous and Exogenous Hyaluronan in Otology,” in 1994 NovelBiomaterials Proceedings, supra, by Davidson et al., “BiologicalResponse of Experimental Cutaneous Wounds in the Pig to Hyaluronan EsterBiomaterials,” loc. cit., by Donati et al., “In Vitro Development andClinical Application in Burn Patients of Keratinocytes Cultured in aHyaluronic Acid Ester Membrane,” loc. cit, and by Benedetti (1994),supra.

The present invention also contemplates the use of chitin, chitosan andderivatives thereof The chitin, chitosan and derivatives thereof may beformed into various materials, such as films, fibers and membranes. Thefibers may be utilized to produce various fabrics, including woven ornonwoven materials. The membranes also can be made into perforatedmembranes. Materials comprised of chitin, chitosan and derivativesthereof are detailed by Rathke and Hudson, in “Review of Chitin andChitosan as Fiber and Film Formers,” Rev. Macromol. Chem. Phys. C34 (3):375-437 (1994), herein incorporated in its entirety by reference.

The fibrinogen component of the compositions can be obtained by anyknown method. Examples of such methods include centrifugation,cryoprecipitation and precipitation using polyethylene glycol, ether,ethanol, glycine or ammonium sulfate. Methods of obtaining suitablefibrinogen are disclosed, for example, by Brennan, “Fibrin Glue,” BloodReviews 5: 240-44 (1991). Lyophilized fibrinogen, which is particularlysuited for use in the present invention, is described in PCT ApplicationWO 9617631 and in U.S. Pat. Nos. 4,909,251 and 4,650,678. Additionalfibrinogen containing products suitable for use in the present inventionare disclosed in U.S. Pat. Nos. 5,290,918 and 5,395,923.

A thrombin-containing component is a common element of conventionalfibrin glues. The thrombin may be derived from any suitable source andobtained by any known method, including recombinant methods. Bovine andhuman derived thrombins are examples of suitable thrombins. The thrombincan be in powder form and can contain additional elements such as CaCl₂.

In addition to or in substitution for thrombin, otherfibrinogen-cleaving substances can be employed in the present invention.Illustrative examples of such fibrinogen-cleaving substances includeenzymes such as those found in snake venom, for example batroxobin,calobin, fibrozyme and enzymes isolated from the venom of Bothropsjararacussu. See Dascombe et al., “Application of Thrombin Based FibrinGlue and Non-Thrombin Based Batroxobin Glue on Intact Human BloodVessels: Evidence for Transmural Thrombin Activity,” Thromb. Haemost.78: 947-51 (1997); Hahn et al., “Purification and Molecular Cloning ofCalobin, a Thrombin-Like Enzyme from Agkistrodon Caliginosus (KoreanViper),” J. Biochem. (Tokyo) 119: 835-43 (1996); Fortova et al.,“Simultaneous Isolation of Protein C Activator, Fibrin Clot PromotingEnzyme (Fibrozyme) and Phospholipase A₂ from the Venom of the SouthernCopperhead Snake,” J. Chromatogr. S. Biomed. Appl. 694: 49-53 (1997);Andriao-Escarso et al., “Isolation and Characterization of a NewClotting Factor from Bothrops Jararacussu (Jararacucu) Venom,” Toxicon.35: 1043-52 (1997).

The coagulation factors and other optional components may be applied tothe hyaluronic acid derivative material either as a dry preparation, anaqueous or nonaqueous preparation or combination thereof. In oneembodiment, the coagulation factors and optional other components are adry preparation and are applied to the HA or HA derivative material byany suitable method which can achieve uniform application. For example,the coagulation factors and optional other components may be sprayedonto the material. The coagulation factors and optional other componentsmay be applied together or sequentially and in any order.

In another embodiment the fibrinogen and thrombin, as well as anyadditional constituents that can be suitably sprayed as an aerosol, canbe sprayed uniformly over the hyaluronic material, thereby beinguniformly applied onto the hyaluronic material.

In yet another embodiment, the coagulation factors and optional othercomponents are chemically bonded to the HA or HA derivative material.

The hyaluronic material, with the fibrinogen, thrombin, and anyadditional components applied thereto may optionally be sealed. Anysuitable, inert coating material can be used to seal the powders ontothe film. Examples of such suitable coatings are vitamin based coatings,such as B12- and riboflavin-based coatings.

In alternative to or in combination with a sealing layer, an additionallayer may be placed on top of the side comprising the coagulationfactors and optional other components. In one embodiment, thisadditional layer comprises an HA or HA derivative material. The materialmay be the same or a different formulation than the material onto whichthe coagulation factors and optional other components are applied orchemically bonded. In another embodiment, the additional layer comprisesan HA or HA derivative material that is substantially porous orconstructed of a loosely woven fabric, thereby allowing easieractivation of the fibrin glue.

The fibrin adhesives and sealants of the present invention have a widerange of medical uses. They are particularly useful where other suturingtechniques, such as stitching or stapling, are unsuitable. They can beplaced directly on a wound site, and are fully reabsorbed into the body.Where there is a significant amount of blood or other fluid at the site,the fluid will activate the interaction of fibrinogen and thrombin.Where the amount of fluid at the wound site is insufficient, a saline orother suitable solution may be used to activate the composition at thewound site. The fibrin adhesives and sealants of the present inventionalso are beneficial in absorbing excess fluid at a wound site. Stillfurther, the compositions of the present invention may serve as asurgical adhesion barrier.

From the foregoing description, one skilled in the art can easilyascertain the essential characteristics of the invention and, withoutdeparting from the spirit and scope of the invention, can make changes,modifications, and variations of the invention to adapt it to varioususes and conditions.

What is claimed is:
 1. A method for preparing a bioabsorbablecomposition, comprising providing a biomaterial which is selected fromthe group consisting of a hyaluronic acid material, a hyaluronic acidderivative material, a chitin material, a chitosan material, a chitinderivative material and a chitosan derivative material, and treating thebiomaterial with fibrinogen and fibrinogen-cleaving agent, wherein thefibrinogen and the fibrinogen-cleaving agent are in a non-activatedstate.
 2. The method of claim 1, wherein the fibrinogen andfibrinogen-cleaving agent are sprayed on the biomaterial.
 3. The methodof claim 1, wherein the fibrinogen and fibrinogen-cleaving agent arechemically bonded to the biomaterial.
 4. The method of claim 1, whereinthe fibrinogen and fibrinogen-cleaving agent are sealed to thebiomaterial.
 5. A bioabsorbable material, comprising a composition whichincludes fibrinogen, a fibrinogen-cleaving agent and a biomaterial whichis a hyaluronic acid material, a hyaluronic acid derivative material, achitin material, a chitosan material, a chitin derivative material and achitosan derivative material, wherein the composition is obtained bytreating the biomaterial with fibrinogen and the fibrinogen-cleavingagent, and the fibrinogen and the fibrinogen-cleaving agent are in anon-activated state.
 6. The material of claim 5, further comprising asealing layer.
 7. The material of claim 5, further comprising anadditional biomatetial layer.
 8. A method of achieving homeostatis,gluing or wound healing, comprising providing a patient having a site ofinjury or incision in need of homeostatis, gluing or wound healing, andapplying a fibrin adhesive or sealant composition comprising fibrinogen,a fibrinogen-cleavirig agent and a biomaterial which is a hyaluronicacid material, a chitin material or a chitosan material, wherein boththe fibrinogen and the fibrinogen-cleaving agent are incorporated on thebiomaterial, to the site.
 9. The method of claim 8, further comprisingadding a saline solution to the composition.
 10. A method of achievinghomeostatis, gluing or wound healing, comprising providing a patienthaving a site of injury or incision in need of homeostatis, gluing orwound healing, and applying a biocompatible and bioabsorbable materialcomprising a biomaterial which is selected from the group consisting ofa hyaluronic acid material, a hyaluronic acid derivative material, achitin material, a chitosan material, a chitin derivative material and achitosan derivative material; fibrinogen; and thrombin; wherein both thefibrinogen and the thrombin are incorporated on the biomaterial to thesite.
 11. The method of claim 10, further comprising adding a salinesolution to the material.